Descarbamoylbluensidine and process for the preparation thereof



United States Patent DESCARBAMOYLBLUENSIDINE AND PROCESS FOR THEPREPARATION THEREOF Alexander D. Argoudelis and Brian Bannister,Kalamazoo,

Mich., assignors to The Upjohn Company, Kalamazoo, 5

Mich, a corporation of Delaware No Drawing. Original application Nov.15, 1962, Ser. No. 238,047, now Patent No. 3,207,780, dated Sept. 21,1965. Divided and this application Aug. 31, 1964, Ser.

5 Claims. (Cl. 260564) This application is a division of applicationSerial No. 238,047, filed on November 15, 1962, and now US. Patent No.3,207,780.

This invention is related to novel compositions of matter and to aprocess for the preparation thereof, and is particularly directed tonovel compounds derived from the antibiotic bluensin and to processesfor producing the same.

More particularly, the invention relates to bluensidine,1-deoxy-1-guanidino-3 -O-carbamoyl-scyllo-inositol, a com- 0 poundhaving the following formula:

and to its preparation.

Still more particularly, the invention is directed to acylates ofbluensidine having the formula:

IfiIR NHC-NHR i O R wherein R is an acyl radical selected from the groupconsisting of hydrocarbon acyl of from 2 to 12 carbon atoms, inclusive,and to their preparation.

Still more particularly, the invention relates todescarbamoylbluensidine, l-deoxy-1-guanidino-scyllo-inositol, a

compound having the formula:

l toand to its preparation.

Still more particularly, the invention relates to acylates ofdescarbarnoylbluensidine having the formula:

(III) tact; ..L. 3.13;?

Patented July 12, 1966 Still more particularly, the invention relates tobluensurea, l-deoxyl-l-ureido-scyllo-inositol, a compound having theformula:

II NH-C-NE:

and to its preparation.

Still more particularly, the invention relates to acylates of bluensureahaving the formula:

(VI) wherein R is as defined above and to its preparation.

The starting material, bluensin, is a bio-synthetic product produced bythe controlled fermentation of Streptomyces bluensis var. bluensis. Thestructure of bluensin is now shown to be (VIIa or VIIb).

HO OH OB I Hsi7\OH H/ H HOHzC H O \l HO CHzOH I l H CHr-NH H \I l/ OH Hstreptobiosaminide are concomitantly formed. The bluen-- sidine can beisolated as its salts by conventional means, for example, adsorptionchromatography.

Bluensidine is a nitrogenous base and as such can exist in both theprotonated and nonprotonated form according to the pH of theenvironment. The protonated form can be isolated as acid addition saltswhich are useful for upgrading the free base, i.e., the nonprotonatedform. Suitable acids for this purpose are hydrochloric acid, sulfuricacid, phosphoric acid, thiocyanic acid, fluosilicic acid, picric acid,Reineckes acid, azobenzenesulfonic acid, and the like.

Bluensidine is a very strong organic base and can be used as an antacidand as the basic catalyst in basecatalyzed organic reactions, e.g., inthe Knoevenagel condensation of aldehydes with reactive methylenegroups. It is also useful as an intermediate. Thus, the condensationproducts obtained from the thiocyanic acid addition salts andformaldehyde according to U.S. Patents 2,425,- 320 and 2,606,155 areuseful as pickling inhibitors, and the fluosilicic acid addition saltsare useful as moth-proofing agents according to U.S. Patents 1,915,334and 2,075,359.

On acylation of bluensidine, the hexaacylate (II) is obtained. Anystandard acylating agent, e.g., acyl halide and anhydrides of theformula R-Hal, R 0, wherein Hal is halogen, for example, chlorine,bromine, fluorine, and iodine, and R is selected from the classconsisting of hydrocarbon carboxylic acid acyl of from 2 to 12 carbonatoms inclusive, can be used.

The term hydrocarbon carboxylic acid acyl of from two to twelve carbonatoms whenever used in the specification or claims is intended to meanan acyl corresponding to a hydrocarbon carboxylic acid of from two totwelve carbon atoms, inclusive. Suitable such acids include (a) asaturated or unsaturated, straight or branched chain aliphaticcarboxylic acid, for example, acetic, propionic, butyric, isobutyric,tert-butylacetic, valeric, isovaleric, caproic, caprylic, decanoic,dodecanoic, acrylic, crotonic, hexynoic, heptynoic, octynoic acids, andthe like; (b) saturated or unsaturated cycloaliphatic carboxylic acid,for example, cyclobutanecarboxylic acid, cyclopentanecarboxylic acid,cyclopentenecarboxylic acid, methylcyclopentenecarboxylic acid,cyclohexanecarboxylic acid, dimethylcyclohexenecarboxylic acid,dipropylcyclohexanecarboxylic acid, and the like; (c) a saturated orunsaturated cycloaliphatic-substituted aliphatic carboxylic acid, forexample, cyclopentaneacetic acid, cyclopentanepropionic acid,cyclopenteneacetic acid, cyclohexanebutyric acid,methylcyclohexaneacetic acid, and the like; (d) an aromatic carboxylicacid, for example, benzoic acid, toluic acid, naphthoic acid,ethylbenzoic acid, isobutylbenzoic acid, methylbutylbenzoic acid, andthe like; and (e) an aromatic aliphatic carboxylic acid, for example,phenylacetic acid, phenylpropionic acid, phenylvaleric acid, cinnamicacid, phenylpropiolie acid, and naphthylacetic acid, and the like.

The hexaacylates of bluensidine are useful as intermediates, forexample, in the preparation of descarbamoylbluensidine and bluensurea.Further, the hexaacylates can be used to upgrade bluensidine; thus, byacylating crude bluensidine, purifying the acylate, and then convertingit back to bluensidine, for example, by ammonolysis, non-acylatable andother impurities are separated.

Descarbamoylbluensidine (III) is obtained by acid hydrolysis ofbluensidine or its hexaacylate. For example, on treating bluensidine orits acylate with dilute aqueous hydrochloric acid,descarbamoylbluensidine is obtained as the hydrochloride which can berecovered by crystallization or other known techniques.

Descarbamoylbluensidine is a nitrogenous base and like bluensidine canexist in both the protonated and nonprotonated form according to the pHof the environment. The protonated form can be isolated as acid additionsalts which are useful for upgrading the free base, i.e., thenon-protonated form. Suitable acids for this purpose are those listedabove.

Further, descarbamoylbluensidine can be used as an antacid and as thebasic catalyst in base-catalyzed organic reactions, e.g., in theKnoevenagel condensation of aldehydes with reactive methylene groups. Itis also useful as an intermediate. Thus, the condensation productsobtained from the. thiocyanic acid addition salts and formaldehydeaccording to U.S. Patents 2,425,320 and 2,606,155

are useful as pickling inhibitors and the fluosilicic acid additionsalts are useful as mothproofing agents according to U.S. Patents1,915,334 and 2,075,359.

On acylation of descarbamoylbluensidine, the heptaacylate (IV) isobtained. Any standard acylating agent, e.g., acyl halides andanhydrides of the formula: R-Hal, R 0, wherein Hal and R are as definedpreviously.

The heptaacylates of descarbamoylbluensidine are useful to upgradedescarbamoylbluensidine; thus, by acylating crudedescarbamoylbluensidine, purifying the acylate, and then converting itback to descarbamoylbluensidine, for example, by ammonolysis,non-acylatable and other impurities are separated.

On reacting bluensidine or its hexaacylate with an alkaline materialthere is obtained bluensurea (V). For example, on reacting bluensidineor hexaacetylbluensidine with saturate-d aqueous barium hydroxide for 1hour there is obtained bluensurea. Other alkaline materials such asalkali metal hydroxides, for example, sodium hydroxide, potassiumhydroxide, lithium hydroxide, and the like, alkaline earth metalhydroxides, for example, calcium hydroxide, and alkali metal carbonates,for example, sodium carbonate, potassium carbonate, and the like can beused. Conversion to bluensurea is also effected when bluensidine or itshexaacylate is brought into contact with or passed over a very stronglybasic anion exchange resin. [Suitable anion exchange resins for thispurpose are obtained by chloromethylating by the procedure given onpages 88 and 97 of Kunin, Ion Exchange Resins, 2nd ed. (1958), JohnWiley & Sons, Inc., polystyrene crosslinked, if desired, withdivinylbenzene prepared by the procedure given on page 84 of Kunin,supra, and quaternizing with trimethylamine or dimethylethanolamine(Dowex 2) by the procedure given on page 97 of Kunin, supra. Anionexchange resins of this type are marketed under the trade names Dowex 2,Dowex 20, Amberlite IRA-400, Duolite A-102, and Permutit 8-1.]

Bluensurea decomposes nitrous acid and, therefore, is useful when anexcess of nitrous acid is to be destroyed in diazotizations. Also,bluensurea is useful as an intermediate. Thus, the condensation productsobtained from the thiocyanic acid addition salts and formaldehydeaccording to the U.S. Patents 2,425,320 and 2,606,155 are useful aspickling inhibitors, and the fluosilicic .acid addition salts are usefulas mothproofing agents according to U.S. Patents 1,915,334 and2,075,359.

0n acylation of bluensurea (V) in the manner described above forbluensidine (I), the pentaacylate (VI) is obtained.

The pentaacylates of bluensurea are useful to upgrade bluensurea; thus,by acylating crude bluensurea, purifying the acylate, and thenconverting it back to bluensurea, for example, by ammonolysis,non-acylatable and other impurities are separated.

On reacting bluensidine (I) or its acylate (H), descarbamoylbluensidine(III) or its acylate (IV), or bluensurea (V) or its acylate (V1), withan alkaline material there is obtained scyllo-inosamine. For example,upon reacting the above named compounds with a saturated aqueous bariumhydroxide solution for 19 hours there is obtained scyllo-inosamine.Other alkaline material listed previously can be used. As is well knownin the art, scyllo-inosamine can be deaminated to myo-inositol byreacting with aqueous nitrous acid. Bluensurea likewise can be convertedto myo-inositol by reacting with nitrous acid. It has been shown thatanimals require myoinositol for the synthesis of phospholipids. [For asummary of the physiological activity of myo-inositol, see S. J. Angyaland L. Anderson, the Cyclitols in Advances in Carbohydrate Chemistry,14, (1959), and references quoted therein] In addition, myo-inositol hasbeen shown to increase the yield of streptomycin when added to growingcultures of Streptomyces griseus [S. K. Majundar and A. I. Kutzner,Science, 135, 734 (1962)].

5 The following examples are illustrative of the process and products ofthe present invention, but are not to be construed as limiting. Allpercentages are by weight and all solid mixture proportions are byvolume unless otherwise noted.

PREPARATION OF BLU'ENSIN Fermentation.A soil stock of Streptomycesbluensis var. bluensis NRRL 2876, was used to inoculate 100 ml. ofsterile medium in a 500 ml. flask. The medium was of the followingcomposition:

The medium was incubated for 2 days at 28 C. on a reciprocating shaker.A culture medium thus obtained was used to inoculate 20 liters ofsterile seed medium contained in a 30 liter stainless steel tank. Themedium was of the following composition.

g. Wilson's Peptone No. 159 10 Corn steep liquor l Refined cottonseedmeal 2 Glucose monohydrate 10 Water to make 1 liter.

(pH adjusted to 7 prior to sterilization) An enzymatic hydrolysate ofproteins of animal origin, protein content approximately 57 percent.

The tank medium was incubated at 28 C. for 1 day and the contents werestirred continually with sparge-d air at the rate of 10 liters of airper minute. Twelve liters of the resulting seed growth medium was usedto inoculate 250 liters of sterile fermentation medium contained in a100 gal-Ion stainless steel fermcnter. The medium was of the followingcomposition:

g. Glucose monohydrate 10 Black strap molasses 30 Brewers yeast Cornsteep liquor 20 Water to make 1 liter.

(pH adjusted to 7.2 prior to sterilization) The fermenter medium wasincubated at 28 C., sparged with air at the rate of 100 liters of airper minute, and agitated by an impeller at 280 r.p.m. The beer washarvested after 88 hours of fermentation.

Isolation.-Sufficient whole beer was produced in the manner as describedabove to yield 5350 liters at pH 8.2. The whole beer was treated with 16kg. of oxalic acid, adjusted to pH 2.9 with l N sulfuric acid, andfiltered using about 160 kg. of diatomite and about 500 liters of washwater. The filtered beer, now about 5400 liters, was adjusted to pH7.8-8.0 with percent aqueous sodium hydroxide and polished by filteringthrough a diatomite precoated filter press. The filtered beer (5300liters) was passed downfiow through two resin columns in series. Eachcolumn was 14 inches in diameter and was charged with 4.5 cubic feet ofAmberlite IRC-SO in the sodium cycle. [A cation exchange resin of thecarboxylic type obtained by the copolymerization of acrylic acid anddivinyl benzene by the procedure given on page 87 of Kunin, Ion ExchangeResins, 2nd ed. (1958), John Wiley & Sons, Inc.] The beer was passedthrough the columns at the rate of about 19 liters per minute. The spentbeer was discarded and the columns were washed with deionized water. Theresin columns were then individually eluted with water acidified to pH1.2-1.5 with 1 N sulfuric acid. Each column was eluted 4 times withliters each time. The lead and trail column eluates were individuallypooled, concentrated to about 200 liters each, and each pool wasadjusted to pH 6.5 with 10 percent aqueous sodium hydroxide. Activatedcarbon was added to each of the eluate concentrates to make a slurry,1200 g. and 850 g. being used for the lead and trail columns,respectively, the amount of carbon being dependent upon the solids inthe concentration (1 g. of carbon for each g. of solids). After mixingeach slurry well, the carbon of each was recovered by filtration andeach cake was washed three times, using about 10 liters of water foreach wash. Each cake was then eluted with about 200 liters each of 15percent aqueous acetone.

The aqueous acetone eluate from the lead column amounted to 187 litersand on concentration and freeze drying yielded 1034 g. of bluensin,Preparation 4A. The aqueous acetone eluate from the trail columnamounted to 190 liters and on concentration and freeze drying yielded777 g. of bluensin, Preparation 4B. Preparations 4A and 4B were combinedwith similar material to produce Preparation 40 which was furtherpurified by chromatography.

Adsorption chromatography A carbon column was prepared in the followingmanner. A mixture of 16.5 kilograms each of activated carbon anddiatomite was wet mixed for 1 hour with liters of deionized wateradjusted to pH 3.5 with 1 N sulfuric acid. The slurry was poured into a14 in. column on top of a layer of sea sand and cotton and packed bynitrogen at a pressure of 10 to 15 pounds. A layer of sea sand was addedto the top of the column. The column was four feet tall and comprised aliquid holdup of 155 liters. The feed was 2707 g. of Preparation 4C. Thefeed was dissolved in 6.6 liters of deionized water and adjusted to pH3.5 with 1 N sulfuric acid. Diatomite was added, suflicient to make athick paste, and this thick paste was placed on top of the column. Thecolumn was washed with liters of deionized water and then eluted with300 liters of 10 percent aqueous acetone. Both wash and elution rateswere about 410 ml. per minute. The following eluate pools were made:

Liters of aqueous acetone eluate Pool Solids, Number mgJml.

1 6 2 5 2a 5 3 7 4 27 5 7 6 2 discarded.

The activity closely follows the solids content of each pool. Thegreatest amount of activity was found in pool No. 4 (K=0.38) which ondrying yielded 640 g. of bluensin, Preparation 5A.

Preparation 5A was reslurried in 670 ml. of deionized water, the mixturewas heated to 45 C. and then filtered with the aid of diatomite. Thefiltrate was diluted with 2.5 liters of water and freeze-dried to form acake. The cake was slurried in 1 liter of deionized water at 35 C. andthen freeze-dried to produce 490 g. of essentially pure bluensinsulfate, Preparation 53.

Characterization Preparation 5B was characterized as follows:

Infrared absorption (in mineral oil mull): The infrared spectrum showscharacteristic absorption, expressed in reciprocal centimeters, at thefollowing frequencies: 3350-2950, 1695, 1655, 1610, 1140-960, 850, 760.

Ultraviolet absorption (in water): The ultraviolet spectrum shows noabsorption maxima between 232 and 400 mp.

7 Elemental analysis.--Found: C, 34.88; H, 5.63; N, 10.07; 0, 45.01; S,4.59.

Paper chromatography: Bluensin shows the following Rf ranges.

Solvent system: Rf range (1) l-butanolzwater (84:16) 0.00.15 (2)l-butanolzwater (84:16) plus 0.25

percent p-toluenesulfonic acid 0.00.15 ('3) 1-butanol:acetic acidzwater(2:1:1) 0.05-0.25 (4)1-butanol1water (84:16) plus 2 percent piperidine0.00.15 (5) l-butanoltwater (96.4) 0.2-0.9 (6) l-butanoltwater (96.4)plus 0.25 percent p-toluenesulfonic acid 0.05-0.25 (7) 1-butanol:water(84:16) plus 2 percent p-toluenesulfonic acid 005-025 (8) Methanolzwatercontaining 3 percent NaCl (80:20) 0.4-0.65

Equivalent weight: 657. pKa': Sulfate salt (in water) 7.6. Specificrotation: [a] =87 (in water). Spot tests: Sakaguchi-positive,Maltolnegative,

Ninhydrinnegative, Biuret--negative, Polypeptide negative,

Slubilz'ty.-BluenSin is soluble in water to the extent of more than 500and less than 1000 mg./ml. The compound is soluble in the followingsolvents to an extent of less than 1 mg./ml.: pyridine, chloroform, 90percent ethanol, ethyl acetate, cyclohexane, benzene, acetone,dirnethylformamide and dioxane.

Purity.-The compound, Preparation B, was found to be 98.2:08 percentpure by phase solubility analysis in a mixture of 70 percent acetone and30 percent water by volume.

PREPARATION OF BLUENSIN DIHYDROCHLORIDE Bluensin sulfate (100 g.) wasdissolved in 800 ml. of water. The pH of the solution was adjusted to4.5 with l N aqueous hydrochloric acid. This solution was passed througha resin column containing one kg. of Dowex 2, an anion exchange resinformed by chloromethylating divinylbenzene (8 percent crosslinked) andquaternizing with dimethylethanolamine. The resin was in the chlorideion form. The column was washed with water. The eluate (1200 ml.) wasfreeze-dried to 84.4 g. of bluensin dihydrochloride.

EXAMPLE 1 Bluensidine A. METHANOLYSIS 0F BLUENSIN HYDROCHLORIDE ANDISOLATION OF BLUENSIDINE CARBONATE Bluensin dihydrochloride (18.3 g.)was dissolved in 600 ml. of 1 N methanolic hydrogen chloride solution.The solution was allowed to stand at room temperature for 48 hours.Silver carbonate (113 g.) was added under vigorous stirring. Insolublesilver carbonate and silver chloride were separated by filtration andthe filtrate was concentrated to dryness. The residue was used for acarbon chromatographic column which was prepared as follows: 200 gramsof activated carbon and 300 grams of diatomite were stirred with 3000ml. of Water. The mixture was poured into a glass column (2 in. insidediameter). The column was packed under pressure of 2 lbs. per sq. inchuntil a constant height was obtained. It was then washed with 2 hold-upvolumes of water (holdup vo1ume=1200 ml.). Sea sand was added on the topof the carbon bed. The residue obtained from the methanolysis, describedabove, was dissolved in 100 ml.

1 Ninhydrln on acid hydrolysate.

of water and then added at the top of the column. The column was theneluted with water and fractions of 20 ml. each were collected. The first59 fractions were discarded. Every fifth fraction was then analyzed bysolids determination with the following results.

Pools of the above fractions were made and freeze-dried to give thefollowing bluensidine carbonate preparations:

Pool 1 Fractions 75-85 (0.94 g.) P0012 Fractions 86-105 (3.59 g.) P0013Fractions 106-124 (0.97 g.)

All three preparations had identical infrared spectra, which showedabsorption bands (in mineral oil mull) at 3450-3300 cm.- (OH, NH), broadband at 2000 cm.- (CO =),1710,1667,1639, 1600 cmr B. METHANOLYSIS OFBLUENSIN DIHYDROCHLORIDE AND ISOLATION OF BLUENSIDINE HYDROCHLORIDEBluensin dihydrochloride g.) was dissolved in 2400 m1. of a methanolichydrochloric acid solution prepared by mixing acetyl chloride andmethanol (792921 v./v.) The solution was allowed to stand at roomtemperature for 48 hrs., and then was mixed with 7200 ml. of ethylether. A precipitate formed and was allowed to settle at roomtemperature for 12 hrs. The supernatant was then decanted. Theprecipitate was washed with ether, isolated by filtration and dried;yield 60 g. This material was used as a feed for a carbonchromatographic column which was prepared as follows: 300 g. ofactivated carbon and 600 g. of diatomite were stirred with 4000 ml. ofwater. The mixture was poured into a glass column (2 in. insidediameter). The column was packed under a pressure of 2 lbs. per sq. in.until a constant height was obtained. It was then washed with 2 holdupvolumes of water. Sea sand was added at the top of the carbon bed. Thefeed material was dissolved in ml. of water and added at the top of thecolumn. The column was then eluted with water and fractions of 20 ml.each were colvlected. The first 100 fractions were discarded. Everyfifth fraction was then analyzed by solids determination with thefollowing results.

Fraction No. Solids (mg/2 ml.)

9 Fraction No. Solids (mg./ 2 ml.)

Pools of the above fractions were freeze-dried to yield the followingbluensidine hydrochloride preparations.

Pool No: Fraction No. 1 120-125 (1.78 g.) 2 126-130 (3.87 g.) 3 131-135(3.10 g.) 4 136-145 (2.80 g.) 5 146-189 (2.55 g.)

The above 5 pools were dissolved in methanol, and the methanolicsolution was added to 300 ml. of ether. Precipitated materials wereseparated by filtration and dried to give bluensidine hydrochloridePreparations 1, 2, 3, 4, and 5 which had the following physical andchemical characteristics.

Infrared spectra: The infirared spectra of Preparations 1 thru 5 werefound to be identical as follows: 3330 (s), 3200 (s), 2940 (oil), 2915(oil), 2845 (oil), 1710 (s), 1670 (s), 1625 (s), 1458 (oil), 1373 (oil),1350 (s), 1215 (m), 1107 (s), 1060 (m), 1033 (m) 1007 (s), 967 (w), 920(w), 775 (m), 718 (oil), cm.-

U.V. spectra: Preparations 1 thru 5 did not show any absorption maximumbetween 220 and 400 mu.

Optical rotation: Prep. No.: [@1 1 +9.0 2 +1.8 3 +1.5 4 +0.4 5 +0.5

[c.=1 (water)] Elemental analysis.-Calculated for C H N O 'HCI: C,31.92; H, 5.69; N, 18.62; Cl, 11.78. Found: Preparation No. 3: C, 31.64;H, 5.81; N, 17.60; Cl, 11.94. Preparation No. 4: C, 32.10; H, 5.67; N,17.36; Cl. 11.63.

EXAMPLE 2 H exaacetylbluensidine bicarbonate, and twice again withwater. Removal of the solvent, after drying over anhydrous sodiumsulfate, on a rotating evaporator at 40/ 15 mm. gave a pale yellowviscous oil (2.58 g.) which crystallized on treatment with ethylacetate; crystals melted at 245-248 C. Recrystallization from ethylacetate-Skellysolve B (isomeric hexanes) gave small colorless prisms ofhexaacetylbluensidine having a M.P. 249-251 C., and 250-251 C. on asecond recrystallization.

Elemental analysis.-Calculated for C H- O N C, 46.51; H, 5.46; N, 10.85;acetyl, 49.99, mol. wt. 516. Found: C, 46.73; H, 5.61; N, 11.13; acetyl,50.15, mol. wt. 494 (osmometric, in CHCl Hexaacetylbluensidine (300 mg.)was added at 0 C. to methanol (100 cc., saturated with dry ammonia at 0C.), and the colorless solution left overnight. Removal of the solventin vacuo gave a colorless mixture of amorphous and crystal-line materialwhich was dissolved in methanolic hydrogen chloride (8 ml. of N). Etherwas added carefully until no further precipitation occurred. Ondecantation, washing with ether, and drying at room temperature, 124 mg.of bluensidine hydrochloride was obtained.

EXAMPLE 3 Descarbamoylbluensidine hydrochloride Bluensidinehydrochloride (one gram) was dissolved in 100 ml. of 1.5 N aqueoushydrochloric acid. The solution was kept under reflux for 15.5 hours; itwas then cooled and concentrated to dryness under reduced pressure. Theresulting residue was dissolved in percent aqueous methanol (10 ml.),and acetone was added until the solution became cloudy. Upon permitting.the solution to stand at room temperature for 2 hrs., crystallinedescarbamoylbluensidine hydrochloride precipitated. The crystals wereisolated by filtration and dried; yield 500 mg. An additional 150 mg. ofcrystals were isolated from the mother liquors. A total of 2.5 mmoles ofguanidinoinositol hydrochloride was isolated from 3.3 mmoles ofbluensidine hydrochloride or 0.76 mole of descarbamoylbluensidinehydrochloride per mole of starting material. Recrystallization of theabove crystals gave descarbamoylbluensidine hydrochloride crystals whichhad the following chemical and physical properties.

Appearance: Colorless needles.

Melting point: 228-230 C. (decomposition).

Optical rotation: [oc] =0 (c.=1, water).

I.R. spectrum: 3435 (s), 3370 (s), 3335 (s), 3270 (s),

. 3180 (s), 2940 (oil), 2918 (oil), 2840 (oil), 1665 (s),

1623 (s), 1573 (w), 1458 (oil), 1373 (oil), 1365 (m), 1347 (W), 1328(m), 1312 (W), 1290 (In), 1273 (m), 1320 (m), 1220 (w), 1167 (W), 1132(In), 1113 (s), 1100 (IS), 1072 (m), 1062 (m), 1045 (w), 1010 (s), 995(s), 963 (w), 708 (m), cm.

Band intensities are indicated as s, m, and w, respectively, and areapproximated in terms of the background in the vicinity of the bands. Ans band is of the same order of intensity as the strongest band in thespectrum; m bands are between one-third and two-thirds as intense as thestrongest band and w bands are less than one-third as intense as thestrongest band. These estimates are made on the basis of a percenttransmission scale.

U.V. spectrum: No maximum between 220-400 m Elementalanalysis.-Calculated for C7H15N3O5'HCII C, 32.59; H, 6.25; N, 16.29; 0,31.01; Cl, 13.74; eq. wt. 257.5. Found: C, 32.92; H, 5.95; N, 15.37; 0,31.53; Cl, 13.99; eq. wt. 260.

On reacting descarbamoylbluensidine with acetic anhydride and pyridineas in Example 2 there is obtained heptaacetyldescarbamoylbluensidine.

EXAMPLE 4 Bluensurea ml. of a A precarbonate was again separated byfiltration.

cipitate of barium carbonate formed almost immediately. The solution waskept under reflux for 1 hour. Excess barium hydroxide was precipitatedas barium carbonate by passing carbon dioxide through the reactionmixture. The precipitated barium carbonate was removed by filtration andthe filtrate was freeze-dried to give 2.95 g. of a colorless amorphousmaterial. Crystallization from 50 percent aqueous methanol-acetoneafforded 1.2 g. of bluensurea as a colorless crystalline material havingthe following physical and chemical properties:

Appearance: Colorless irregular plates.

Melting point: 244-245 C. with decomposition.

Optical rotation: [a] (c.=1, water) Infrared spectrum: 3460 (s), 3390(s), 3360 (s), 3310 (s), 3190 (s), 2940 (oil), 2920 (oil), 2845 (oil),1680 (s), 1640 (m), 1620 (s), 1597 (m), 1550 (s), 1480 (m), 1460 (oil),1375 (oil), 1327 (m), 1277 (m), 1255 (w), 1235 (w), 1218 (w), 1170 (m)1123 (m), 1097 (s), 1055 (m), 1007 (s), 995 (s), 773 (w), cm.-

UV. spectrum: No maximum between 220-400 m Elemental almIysis.Calculatedfor C- H N O C, 37.87; H, 6.36; N, 12.62; 0, 43.24. Found: C, 37.90; H,6.21; N, 11.96; 0, 43.99.

Potentiometric titration: No titrable group.

On reacting bluensurea with acetic anhydride and pyridine as in Example2 there is obtained pentaacetylbluensurea.

EXAMPLE Preparation of scyllo-inosamine hydrochloride Bluensidinehydrochloride (3 grams) was dissolved in 300 ml. of saturated aqueousbarium hydroxide solution. The solution was kept under reflux for 19hours after which it was cooled. Precipitated barium carbonate wasseparated by filtration. The filtrate was saturated with carbon dioxideand the precipitated barium The filtrate was freeze-dried to give 2.43g. of a colorless amorphous solid. 500 mg. of this material wasdissolved in 50 ml. of a solution of 0.2 N methanolic hydrochloric acidand 5 ml. of water. Upon the addition of acetone, colorlessscyllo-inosamine hydrochloride crystals precipitated. These crystalswere filtered off, washed with acetone and dried; yield 375 mg.

Recrystallization of these crystals from water-ethanolacetone (1:1:1)gave 60 mg. of colorless irregular plates which darken at approximately260 C. and do not melt below 300 C. and having the following:

Elemental analysis.--Calculated for C H NO Cl: C, 33.52; H, 6.56; N,6.52; Cl, 16.42. Found: C, 33.54; H, 6.65; N, 6.81; Cl, 15.21.

EXAMPLE 6 Preparation of myo-inositol lzexaacetate Scyllo-inosaminehydrochloride (700 mg.) was dissolved in 10.5 ml. of water and 1.4 ml.of acetic acid. This solution was mixed with a solution of 1.75 g. ofbarium nitrite in 8.75 ml. of water. The mixture was allowed to stand at5 C. for 21 hours. Sulfuric acid (2 N) was then added, and theprecipitated barium sulfate was separated by filtration. The filtratewas concentrated in vacuo to a volume of 5 ml. myo-inositol as acolorless crystalline material, then precipitated. The crystals weredissolved in 30 ml. acetic anhydride, mixed with 50 mg. of sodiumacetate and the mixture was kept under reflux for 30 min. The mixturewas added to 400 ml. of water, and the aqueous solution was extracted 3times with 200 ml. of chloroform. Concentration of the chloroformextract afforded myo-inositol hexaacetate as a crystalline materialwhich was recrystallized from ethanol-water, and which had a meltingpoint of 215-218 C. (reported for myo-inositol hexaacetate 216-217 C.).

EXAMPLE 7 By substituting the acetic anhydride in Example 2 bypropionic, succinic, maleic, and phthalic anhydride, there are obtainedthe corresponding hexapropionyl-, hexasuccinyl-, hexamaleyl-, andhexaphthaloylbluensidine.

EXAMPLE 9 By substituting the bluensidine hydrochloride in Example 5 byhexaacylbluensidine hydrochloride there is obtained scyllo-inosaminehydrochloride.

EXAMPLE 10 By substituting the bluensidine hydrochloride in Example 5 bydescarbamoylbluensidine or heptaacyldesc'arbamoylbluensidinehydrochloride there is obtained scylloinosamine hydrochloride.

EXAMPLE 11 By substituting the scyllo-inosamine hydrochloride in Example6 by bluensurea there is obtained myo-inositol hexaacetate.

We claim:

1. Descar-bamoylbluensidine, a compound which has the structuralformula:

or its acid addition salts.

2. Descarbamoyl-bluensidine.

3. Acid addition salts of descarbamoylbluensidine.

4. A process for making the compound of the formula in claim 1 whichcomprises reacting bluensidine with a dilute aqueous mineral acid underreflux and isolating descarbamoylbluensidine so produced.

5. A process for making the compound of the formula in claim 1 whichcomprises reacting bluensidine with 1.5 N aqueous hydrochloric acidunder reflux to produce descarbamoylbluensidine.

References Cited by the Examiner UNITED STATES PATENTS 6/1951 Peck260-564 X OTHER REFERENCES Bonnard et al.: Can. J. Chem, vol. 36, pp.1541-1549 (1958).

Rodd: Chemistry of Carbon Compounds, volume 1B, p. 901 (1952).

Taylor et al.: "Sidgwicks Organic Chemistry of Nitrogen, page 273(1945).

CHARLES B. PARKER, Primary Examiner.

JOSEPH P. BRUST, Examiner.

FLOYD D. HIGEL, Assistant Examiner.

1. DESCARBAMOYLBLUENSIDINE, A COMPOUND WHICH HAS THE STRUCTURAL FORMULA: